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direct / indirect detection in WB - (Oct/04/2006 )


For some purposes, sometimes I do a fluorescence WB with a secondary Ab labelled with FITC. But what are the advantages / disadvantages (if they exist) of doing this versus using the primary Ab directly labelled with FITC? unsure.gif

thanks a lot rolleyes.gif


you amplify the signal due to multiplicity of binding sites for the 2nd antibody on the 1st antibody.


mdfenko is right, moreover it is known that secondary and third antibodies (as in ELISA) rise specificity of signal (better signal/noise ratio)

-The Bearer-

Thanks a lot guys, smile.gif smile.gif smile.gif
now I think I´ve heard that explanation before when I was at University and somebody told us something like that.